Functional validation of mungbean LEA protein coding gene in bacterial expression system confers salt stress tolerance

Authors

  • Rajesh SUBRAMANIAN Tamil Nadu Agricultural University, Centre for Plant Molecular Biology and Biotechnology, Coimbatore 641 003, Tamil Nadu; Tamil Nadu Agricultural University, Horticultural College and Research Institute, Periyakulam 625 604, Tamil Nadu (IN)
  • Nandhini U. PANDI Tamil Nadu Agricultural University, Centre for Plant Molecular Biology and Biotechnology, Coimbatore 641 003, Tamil Nadu (IN)
  • Radhamani THANGAVEL Tamil Nadu Agricultural University, Centre for Plant Molecular Biology and Biotechnology, Coimbatore 641 003, Tamil Nadu (IN)
  • Likhith R.K. SWAMY Tamil Nadu Agricultural University, Centre for Plant Molecular Biology and Biotechnology, Coimbatore 641 003, Tamil Nadu (IN)
  • Srimathi Priya LAKSHMINARAYANAN Tamil Nadu Agricultural University, Horticultural College and Research Institute, Periyakulam 625 604, Tamil Nadu (IN)
  • Shenbagavalli SANTHAMANI Tamil Nadu Agricultural University, Horticultural College and Research Institute, Periyakulam 625 604, Tamil Nadu (IN)
  • Backiyavathy M. RAVALAN Tamil Nadu Agricultural University, Horticultural College and Research Institute, Periyakulam 625 604, Tamil Nadu (IN)
  • Rajangam JACOB Tamil Nadu Agricultural University, Horticultural College and Research Institute, Periyakulam 625 604, Tamil Nadu (IN)

DOI:

https://doi.org/10.15835/nbha52313416

Keywords:

characterization, E. coli, LEA, protein expression, salinity, plant stress

Abstract

Mungbean (Vigna radiata R. Wilczek) is a major tropical food grain legume that is widely cultivated in tropical part of the world. Mungbean like other plants, tolerate and survive limited water situation owing to expression of stress associated proteins that offers membrane stability and cell protection. Late Embryogenesis Abundant (LEA) proteins are among the group of low molecular weight proteins, that play diverse roles in stress protection in several species of plants and animals. A LEA protein coding gene VrLEA2 was isolated from mungbean and its role in stress tolerance has been demonstrated using a bacterial expression system. VrLEA2 gene isolated was of size 893 bp and characterized as a group 1 LEA protein based on the sequence signature motif with presence of hydrophilic domain and a characteristic 20-mer conserved amino acids motif. VrLEA2 gene was cloned into a bacterial expression vector, pET 28a (+), transformed into the E.coli BL21 (DE3) cells for recombinant protein expression and subsequently subjected to antibiotic selection with kanamycin. Functional validation of the VrLEA2 for salt stress tolerance with varied concentration of NaCl (0 mM to 600 mM) showed alteration in colony morphology and reduction in the number of colonies in control compared to the transformed cells demonstrating the improved survival rate of cells expressing VrLEA2 protein. These findings indicate the best use of bacterial expression system for functional validation of plant proteins under stressed environments. 

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2024-09-06

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SUBRAMANIAN, R., PANDI, N. U., THANGAVEL, R., SWAMY, L. R., LAKSHMINARAYANAN, S. P., SANTHAMANI, S., M. RAVALAN, B., & JACOB, R. (2024). Functional validation of mungbean LEA protein coding gene in bacterial expression system confers salt stress tolerance. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 52(3), 13416. https://doi.org/10.15835/nbha52313416

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Vol. 52 No. 3 (2024)

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Research Articles
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DOI: 10.15835/nbha52313416

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Copyright (c) 2024 RAJESH S, Nandhini U. PANDI, Radhamani THANGAVEL, Likhith R. K. SWAMY, Srimathi Priya LAKSHMINARAYANAN, Shenbagavalli SANTHAMANI, Backiyavathy M. RAVALAN, Rajangam JACOB

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